Liquid processing method making use of pipette device and apparatus for same

ABSTRACT

Technological preposition of the present invention is a liquid processing method making use of a pipette device which sucks a liquid containing a target high molecular substance from inside of a vessel through a chip detachably set on a sucking port or a discharging port of a liquid sucking/discharging line and transfers this liquid or target high molecular substance to the next target processing position for the purpose to execute such works as quantifying, separating, taking out, pipetting, cleaning, condensing, and diluting a liquid or a target high molecular substance contained in a liquid and also such works as extracting, recovering, and isolating the target high molecular substance by means of sucking and discharging a liquid with a pipette device and controls by a magnetic body over magnetic particles and/or a filter combined according to the necessity, and the chip can isolate the target high molecular substance by having the substance attracted onto magnetic particles or with a filter set on each chip.

TECHNICAL FIELD

[0001] The present invention relates to a liquid processing methodmaking use of a pipette device and an apparatus for the same, with whichworks including for quantifying, separating, taking out, pipetting,clearing, condensing, diluting a liquid or a target high molecularsubstances included in a liquid such as useful substances such asantibiotic substance, genetic substances such as DNA, and immunologicalsubstances such as antibodies, and/or works for extracting, recoveringand isolating the target high molecular substance can automatically andaccurately be executed by means of absorbing and discharging the liquidthrough a liquid absorbing/discharging line in the pipette device.

BACKGROUND ART

[0002] In recent years, research activities for DNA or the like are veryactive in many fields including engineering, medical science,agriculture, physical science, pharmacology, and the purpose includesgenome sequencing, clinical diagnosis, improvement of agriculturalproducts, bacteriological inspection of foods, drug preparing systems orthe like.

[0003] As described above, when various types of immunoassay applicablein a very wide range with high expected possibility in its applicationor structural analysis of molecular level organisms, microorganisms, orsubstances such as cells, DNA, RNA, mRNA, plasmid, virus, or bacteria(described simply as a target high molecular substance in the presentspecification) is performed, it is necessary to carry out with highprecision works such as those for quantifying, separating, taking out,pipetting, clearing, condensing, diluting a sample or a target highmolecular substance included in the sample or works for extracting,recovering, or isolating a target high-molecular substance as apreprocessing.

[0004] To explain structural analysis of a gene such as DNA diagnosis asan example, at first it is necessary to extract, recover, and isolate aDNA region including a target gene. The technology for extracting,recovering, and isolating genes has already been established as the genecloning technology or genome sequencing technology, and at present it isbelieved that, by spending enough time and expenses, any gene can beseparated and obtained. For this reason, if a target gene DNA has beenextracted, recovered, and separated, any type of gene analysis ispossible as a principle by making use of the separated gene DNA.

[0005] However, in a case of man, for instance, a particular target geneDNA is one millionth or below of all genome DNA, and for this reasonactually a quantity of DNA obtainable for testing is quite small, whilea quantity of DNA and RNA not necessary for a particular experiment isquite large, which makes it difficult to execute analysis smoothly.

[0006] For the reasons as described above, to execute structuralanalysis of a gene such as DNA diagnosis, it is important to extract,recover, and isolate a DNA area including a target gene. Description ismade hereinafter for a basic method of extracting, recovering, andisolating a DNA.

[0007] A DNA exist in a nucleus as a complex with a protein in a cell.In the basic sequence for extracting a DNA, a cell or a cell nucleus isprocessed with SDS (surfactant dodecil sodium sulfate) to make the DNAsoluble, and proteins contained therein are removed with a proteolyticenzyme or phenol.

[0008] In other words, when a DNA is separated from the tissue, at firstthe tissue taken out is put in ice and kept therein for a certain periodof time under a low temperature, then this cooled tissue is divided tosmall pieces each with the weight of around 0.1 g, which are washed witha ice-cooled buffer solution A (0.01 M Tris HCl, pH 7.8, 0.1 M NaCl, 2mM MgCl₂) This tissue is put in the above-described buffer solution Ahaving a volume 20 times larger as compared to that of the tissue and ishomogenized 5 or 10 times with a Potter type homogenizer. Then thetissue is put in a centrifugal tube together with the buffer solutionand is subjected to centrifugation (2,000 rpm, for 5 minutes). The cellnucleus or the cell precipitates, so that the supernatant is aborted.When extracting a DNA from cultured cells, the cells are well suspendedin an ice-cooled buffer solution B (0.01 M Tris HCl, pH 7.8, 0.1 M NaCl,2 mM EDTA) and is subjected to centrifugation. The precipitated nucleior cells are again well suspended in the buffer solution B having avolume 100 times larger than that of the specimen.

[0009] After the cells or cell nuclei are well suspended until a blockof cells disappears, a 10% SDS solution is added by one twentieththereof to lyse the cells. Then proteinase K (10 mg/ml) is added by onefiftieth thereof to the solution and reacted for 4 hours under atemperature of 50° C. so that the protein is lysed. During thisreaction, the solution is sometimes agitated because the viscosity ishigh. Then phenol extraction is executed 3 times. In this step, theextracting work should be performed carefully so that no physical poweris not loaded thereto.

[0010] Then the specimen is dialyzed for around 18 hours with a buffersolution C (10 mM Tris HCl, pH 7.8/0.1 mM EDTA) having a volume 100times larger as compared to that of the specimen, and is kept under atemperature of 4° C.

[0011] Through the steps as described above, about 0.2 mg of DNA can beobtained from 0.1 g of tissue. What is described above is a process ofextracting DNA from tissue or cells, and in addition there have beenknown a method in which plasmid DNA is obtained by way of the alkalimethod (the small quantity adjusting method), a method of recovering DNAby way of the boiling method, and a method of recovering closed-cyclebromide DNA by way of the large quantity adjusting method.

[0012] As described above, it is possible to extract, recover, andisolate DNA for structural analysis of a gene in, for instance, DNAdiagnosis according to any of the known methods as described above, buta work for isolating DNA from the tissue or cells as described above is,as clearly understood from the sequence for extracting DNA from thetissue or cells as described above, extremely complicated, and a longperiod of time is required, which is disadvantageous.

[0013] In addition, any of various types of method including thecentrifuging method, high speed liquid chromatography method, gelelectrophoresis method, dispo-column method, dialysis method, glasspowder method, magnetic particle cleaning nozzle method has beenemployed for structural analysis of DNA or the like extracted by theabove-described means, and each of the methods has respective advantagesand disadvantages, and at present a high precision and stable method forstructural analysis has not been developed yet.

[0014] Namely, in a case of centrifugation, automation of processes forloading and taking out vessels is very difficult, and also it is verydifficult to mechanically separate supernatant from precipitates aftercentrifugation, and for this reason its applicability for variouspurpose is disadvantageously poor.

[0015] In a case of high speed liquid chromatography, a separationcolumn is basically consumable, injection for a sample to the column ortime management for separation can not be mechanized, and also differentsamples pass through the column, which disadvantageously makes itimpossible to completely prevent contamination of the column.

[0016] Furthermore, in a case of gel electrophoresis, adjustment of gelcan not be mechanized, and this method has generally been used as abasic technique for separation of DNA, but the separated pieces must betaken out manually, which is disadvantageous.

[0017] The dispo-column method is one of technic which can be embodiedas a kit for separation of a particular DNA piece, but the cost is veryexpensive, and its applicability is narrow. In addition, controls overpipetting and liquid passing through the column are difficult, and thereare many problems in mechanization of this method.

[0018] In the dialysis method, a long period of time is required fordialysis, and also it is hard to apply this method when a quantity ofsample is small, so that this method has not been used widely.

[0019] The glass powder method is an excellent method of extracting DNAmaking use of silicon dioxide, and the process is simple and convenient,but as the powder is separated with a filter or by way ofcentrifugation, it is difficult to automate the entire process.

[0020] Furthermore in a case of the magnetic particle cleaning nozzlemethod, the process can be automated by controlling the cylinder andattracting/discharging with magnetic particles, but basically it isimpossible to prevent contamination only by cleaning the nozzle.

[0021] The present invention was made under the circumstances asdescribed above, and its object is to provide a liquid processing methodas well as an apparatus for the same making use of a completely novelpipette device which can automatically and with high precision executeworks of quantifying, separating, taking out, pipetting, cleaning,condensing, diluting a liquid or a target high molecular substancecontained in a liquid as well as works of extracting, recovering, andisolating the substance by controlling the pipette device's operationsfor sucking or discharging a liquid and magnetic particles with amagnetic body and/or by a combination of a magnetic body and a filter.

DISCLOSURE OF INVENTION

[0022] Technological basis of the present invention is a liquidprocessing method making use of a pipette device which sucks a liquidcontaining a target high molecular substance via a chip detachably setin a sucking port or a discharging port of a liquid sucking/dischargingline from inside of a vessel and transfers the liquid or the target highmolecular substance to a target next processing position, and the chiphas the sucked target high molecular substance deposited on magneticparticles and/or separated with a filter set in the chip. Namely, it ispossible to automatically execute with high precision the works ofquantifying, separating, taking out, pipetting, cleaning, condensing,diluting a liquid or a target high molecular substance as well as worksof extracting, recovering, and isolating the substance by controllingthe pipette device's operations for sucking and discharging the liquidand magnetic particles with a magnetic body and/or by a combination of amagnetic body and a filter.

[0023] Also in the present invention, the target high molecularsubstance is a useful substance such as antibiotics, genetic substancessuch ad DNA, or an immunological substance such as antibody. For thisreason, the present invention is well suited to works of separating,taking out, pipetting, cleaning, condensing, diluting and/or works ofcapturing, extracting, isolating, amplifying, labelling, and measuringmolecule level organisms or microorganisms such as cells, DNA, RNA,mRNA, plasmid, virus, and bacteria or certain high molecular substance,and a target high molecular substance can be obtained without dependingon the conventional centrifugation.

[0024] Also in the present invention, such works as quantifying,separating, taking out, pipetting, cleaning, condensing, and dilutingthe target high molecular substance as described above is carried outwith a chip set in the liquid sucking/discharging line described aboveand at least one type of filter set in a tip section of the chip. Withthis configuration, such works as quantifying, separating, taking out,pipetting, cleaning, condensing, and diluting the target high molecularsubstance can easily be executed with high precision.

[0025] The present invention is embodied mainly as described above, butby providing a plurality of filter holders in multiple stages in a waywhere, for instance, a filter holder with a filter to screen out bloodcorpuscle shells is provided in the first stage of the chip and a filterholder with a silica membrane filter to capture DNA is provided in thesecond stage thereof, such works as quantifying, separating, taking out,pipetting, cleaning, condensing, and diluting the target high molecularsubstance as described above can more easily be carried out with highprecision. It is needless to say that, in a case where separation of atarget high molecular substance is executed by setting each filterholder according to the present invention, the chips and filter holdersmay be engaged in and processed one by one or complied in a multiplestages and set to execute a plurality of works simultaneously.

[0026] Also in the present invention, by using a plurality of filterseach having a different pore size (transmission diameter of each filter)and used for separation of a target high molecular substance and foreignmaterials other than the target high molecular substance respectively,it is possible to obtain only the target high molecular substancewithout fail.

[0027] Also in the present invention, after such works as quantifying,separating, taking out, pipetting, cleaning, condensing, diluting aliquid or a target high molecular substance with the filter as describedabove, in a step of detachably setting a new chip in a tip section ofthe liquid sucking/discharging line and sucking/discharging a solutioncontaining magnetic particles with this chip, as the magnetic particlesare attracted by a magnetic body provided in the side of the chip ontoan internal surface of the chip to extract, recover, and isolate thetarget high molecular substance, such works as quantifying, separating,taking out, pipetting, cleaning, condensing, and diluting and also suchworks as extracting, recovering, and isolating a target high molecularsubstance can automatically be executed.

[0028] Also in the present invention, different from a liquid processingbased on the filter system as described above, such works as capturing,extracting, isolating, amplifying, labelling, and measuring the targethigh molecular substance may be executed only with a chip set in theliquid sucking/discharging line, a magnetic force, and one or aplurality types of magnetic particles and without using the filter asdescribed above, more precise liquid processing can be realized withsimpler configuration.

[0029] Also in the present invention, by causing the chip set in theliquid sucking/discharging line as described above to react withmagnetic particles like in the invention described above, refiningprocess such as capturing cells, having cell cores or protein lysed canautomatically be executed, and a particular target high molecularsubstance can easily be extracted, recovered, and isolated.

[0030] Furthermore in the present invention, by using magnetic particleswith a probe or biotin or streptoavidin coated thereon by making use ofa chip set in a liquid sucking/discharging line like in the inventiondescribed above after the work of extracting the substance, a particularbase sequence piece can easily be isolated with high precision withoutexecuting centrifugation.

[0031] Also in the present invention, it is possible to execute a seriesof works for refining such as capturing cells, or having cell core orprotein lysed by causing a chip set in the liquid sucking/dischargingline to react with magnetic particles for extracting a particular highmolecular substance and then isolating the particular base sequencepiece with other type of magnetic particles with a probe or biotin orstreptoavidin coated thereon easily in a liquid sucking/discharging linein a pipette device.

[0032] Also in the present invention, after a series of works such ascapturing, extracting, and isolating a target high molecular substanceby using the magnetic particles as described above, by making theisolated particular base sequence piece emit light through chemicalluminescence or fluorescence or enzymatic coloration, presence or aquantity of the particular base sequence piece can easily be detected ormeasured.

[0033] Also in the present invention, it is possible to easily andautomatically execute a series of works for refining such as capturingcells, or having cell core or protein lysed by causing a chip set in theliquid sucking/discharging line to react with magnetic particles forextracting a particular high molecular substance, then amplifying theextracted target high molecular substance, isolating the particular basesequence piece with other type of magnetic particles with a probe orbiotin or streptoavidin coated thereon and then detecting presence of ormeasuring a quantity of the particular base sequence piece by causingthe isolated piece to emit light through chemical luminescence orfluorescence or enzymatic coloration.

[0034] In the present invention, by executing the works of separating,taking out, pipetting, cleaning, condensing, diluting the target highmolecular substance and/or works for capturing, extracting, isolating,amplifying, labelling, and measuring the substance in a single liquidsucking/discharging line or a plurality of liquid sucking/discharginglines provided in parallel to each other, a sequences of works can beexecuted efficiently and automatically. In a case where a plurality ofliquid sucking/discharging lines are provided in parallel, theprocessing capacity is improved, and also a multi-channel processingline can be realized.

[0035] In the present invention, in a case where processing is executedwith a plurality of liquid sucking/discharging lines provided inparallel, as the plurality of liquid sucking/discharging lines aredriven and controlled so that the works of separating, taking out,pipetting, cleaning, condensing, diluting the target high molecularsubstance and/or the works of capturing, extracting, isolating,amplifying, labelling, and measuring the substance in each line areexecuted according to the same timing, or also so that each liquid issucked or discharged in a specified processing step according to adifferent timing, and for this reason processing steps suited to atarget high molecular substance can easily be built up.

[0036] In the present invention, by providing working spaces separatedfrom each other with partitions in the single or a plurality of liquidsucking/discharging lines, or by providing working spaces with an airflow by means of continuously sucking air in each line working spacefrom an air sucking port, or by combining these different types ofconfiguration, even in a case of liquid processing such as extractingand analyzing DNA or the like in which it is required to strictlyprevent contamination by air in each processing line, the objective caneasily be achieved.

[0037] Also in the present invention, by having a target high molecularsubstance or a substance bonded to a target high molecular substanceabsorbed or bonded to a surface of each magnetic particle used for thepurpose of the present invention, the target high molecular substancecan be obtained without executing centrifugation.

[0038] In the present invention, in a case where the above-describedmagnetic particles are used, controls are provided so that the magneticparticles are absorbed onto an internal wall of a chip due to a magneticforce working from outside of the chip, or so that, if effect of themagnetic force is weak or not present, the magnetic particles are heldseparable from the internal surface of the chip, it is possible tocontrol capture of target high molecular substance and separation of thesame from foreign materials with high precision.

[0039] In the present invention, by controlling load of a magnetic forceinto or elimination of a magnetic force in a chip by means of moving apermanent magnet in a direction perpendicular to the longitudinaldirection of the chip or in a range including the directionperpendicular to the longitudinal direction of the chip, or by turningON or OFF an electric magnet, it is possible to efficiently execute suchworks as absorbing magnetic particles or agitating and mixing themagnetic particles with other liquid or cleaning.

[0040] In the present invention, in a case where processing is executedwith an electric magnet, by turning ON the electric magnet when itcontacts an external surface of the chip to generate a magnetic force orby moving off the electric magnet from the chip when the magnetic forceis eliminated, absorption of magnetic particles or agitation and mixingof magnetic particles with other solution or cleaning can efficiently beexecuted.

[0041] In the present invention, when removing a chip from a liquidsucking/discharging line, it can easily be removed by holding AS thechip with a holding body synchronously operating when the permanentmagnet or electric magnet moves to the chip and the permanent magnet orelectric magnet and then moving the liquid sucking/discharging lineupward.

[0042] In the present invention, the chip described above comprises asmall diameter section steeped into a liquid, a large diameter sectionhaving a capacity larger than a capacity of a vessel in which a liquidis accommodated, and an intermediate section provided between the smalldiameter section and the large diameter section and having a diametersmaller at least than that of the large diameter section, and magneticparticles are captured by the intermediate section, so that cloggingnever occurs and the magnetic particles can almost completely beabsorbed because of magnetism within a short period of time.

[0043] In the present invention, an internal diameter of theintermediate section of a chip described above has a dimensionappropriate for ferromagnetic field of the magnetic body to provideeffects therein, and magnetic particles are captured due to a magneticforce generated in the ferromagnetic field of the magnetic body, so thatmagnetic particles can almost completely be captured because ofmagnetism within a short period of time.

[0044] In the present invention, by forming an internal diameter of theintermediate section of a chip described above so that it has thesubstantially same width dimension as that of a contacting surface of amagnetic body contacting the intermediate section, the most effectiveabsorption of magnetic particles can be realized.

[0045] In the present invention, absorption of magnetic particles ontoan internal surface of a chip set in the liquid sucking/discharging lineis executed by passing a solution containing magnetic particles througha ferromagnetic field inside a chip at a slow speed appropriate for themagnetic particles to be captured completely, the magnetic particles cancompletely be captured.

[0046] In the present invention, controls are provided so that the finalliquid surface of a liquid passing through the chip when sucked into ordischarged from the chip always reaches the magnetic field describedabove, so that the magnetic particles can be captured more completely.

[0047] In the present invention, in a case where a liquid is sucked ordischarged with a so-called single nozzle, the liquid is sucked bycontacting a tip section for a chip set in the liquidsucking/discharging line to an internal bottom of a vessel with theliquid accommodated therein and then lightly raising the tip section, sothat almost all of the liquid within the vessel can be sucked anduniformity of reaction can be maintained.

[0048] In the present invention, agitation and mixing of magneticparticles absorbed in the chip with a reagent or cleaning water isexecuted under the so-called pumping control in which the works ofsucking and discharging the liquid in the liquid sucking/dischargingline is continuously executed at a high speed and times enough toagitate and mix the liquid with the magnetic particles, so that theliquid and magnetic particles can homogeneously be agitated and mixedwith each other.

[0049] In the present invention, in a case where the works of agitatingand mixing a liquid and magnetic particles with each other are executed,the works of sucking and discharging the liquid in the liquidsucking/discharging line is executed with the tip section steeped into areagent or cleaning water accommodated in a vessel so that a quantity ofa liquid in the vessel and a sucking/discharging rate substantiallycoincide with each other, and for this reason no bubble is generated andthe reaction can be executed under no physical impact, so thatseparation of a target high molecular substance from the magneticparticles due to bubbles can be prevented without fail.

[0050] In the present invention, in a case where necessary temperaturecontrols are provided to promote a reaction between a target highmolecular substance and a reagent or the like or amplification of thetarget high molecular substance, the reaction liquid or a liquid to beamplified is transferred with the chip into each thermostatic chamberpreviously kept in a specified temperature to be heated or cooledtherein, so that a period of time required for heating or cooling thereaction liquid or the liquid to be amplified can substantially bereduced.

[0051] In the present invention, when controlling a temperature in eachreaction chamber, a covering body is set over a tip section of theliquid sucking/discharging line, and the covering body is set via theliquid sucking/discharging line on the thermostatic chamber, so thatevaporation of the liquid can be prevented and also contamination of aircan be prevented without fail.

[0052] In the present invention, the covering body is built so that itis plucked and broken by the liquid sucking/discharging line or a chipset in the line, and for this reason it is not necessary to provide aseparate sucking/discharging means, and a reaction liquid or a liquid tobe amplified in a thermostatic vessel can be sucked by the liquidsucking/discharging line or a chip set therein, so that theconfiguration is quite simple and a series of works can automatically beexecuted.

[0053] In the present invention, to realize each of the liquidprocessing methods as described above, there is provided a liquidprocessing apparatus making use of a pipette device comprising a liquidsucking/discharging line which can move in the horizontal direction andis maintained at a specified position so that it can move in thevertical direction, a means for executing liquid sucking/dischargingworks through the liquid sucking/discharging line, a plurality of chipsrequired for processing one type of liquid and provided along thehorizontal direction in which the liquid sucking/discharging line moves,a vessel with the liquid accommodated therein, one or more filterholders each having a filter required for the processing above, and oneor more vessels each having other type of liquid required for theprocessing above, and the liquid sucking/discharging line or a chip settherein is driven and controlled according to instructions from acontrol unit so that the chip is transferred with a filter holder settherein to execute such works as quantifying, separating, taking out,pipetting, cleaning, condensing, and diluting the liquid or a targethigh molecular substance contained in the liquid, and for this reason itis not necessary to especially provide a means in which an operation maybe interrupted such as a centrifugal separator, and it is possible toautomate such works as quantifying, separating, taking out, pipetting,cleaning, condensing, diluting a target high molecular substance withsimple configuration.

[0054] In the present invention, there is provided a liquid processingapparatus comprising a liquid sucking/discharging line which can move inthe horizontal direction and is maintained at a specified position sothat it can move in the vertical direction, a means for executing liquidsucking/discharging works through the liquid sucking/discharging line, aplurality of chips required for processing one type of liquid andprovided along the horizontal direction in which the liquidsucking/discharging line moves, a vessel with the liquid accommodatedtherein, a magnetic body for attracting magnetic particles contained ina liquid onto an internal surface of a chip when the liquid is suckedinto or discharged from the chip, one or more vessels with other typesof liquid accommodated therein respectively required for the processingdescribed above, and the liquid sucking/discharging line or the chip isdriven and controlled according to instructions from a control unit sothat the chip is transferred to execute such works as capturing,extracting, isolating, amplifying, labelling, and measuring a liquid ora target high molecular substance contained in the liquid, and for thisreason it is not necessary to especially provide a means in which theoperation such as a centrifugal separator may be interrupted, and alsoit is possible to automate such works as capturing, extracting,isolating, amplifying, labelling, and measuring a target high molecularsubstance with simple configuration.

[0055] In the present invention, there is provided a liquid processingapparatus making use of a pipette device comprising a liquidsucking/discharging line which can move in the horizontal line and ismaintained at a specified position so that it can move in the verticaldirection, a plurality of chips required for processing one type ofliquid and provided along the horizontal line in which thissucking/discharging line moves, a vessel with the liquid accommodatedtherein, one or more filter holders each having a filter set thereinrequired for the processing described above, one or more vessels eachaccommodating therein other types of liquid required for the processingabove, a vessel in which a liquid containing magnetic particles isaccommodated, and a magnetic body for attracting the magnetic particlesonto an internal surface of the chip in the process of sucking ordischarging a solution containing the magnetic particles, and the liquidsucking/discharging line is transferred according to instructions from acontrol unit to execute required processing for a liquid or a targethigh molecular substance contained in the liquid, and for this reason itis possible to execute such works as quantifying, separating, takingout, pipetting, cleaning, condensing, diluting a target high molecularsubstance and also such complicated works as extracting, recovering, andisolating the target high molecular substance with very simpleconfiguration in succession and automatically.

[0056] In the present invention, a hook for locking and supporting achip engaged in and supported by the liquid sucking/discharging line isrotatably supported by the liquid sucking/discharging line, and the hookis energized in its normal state in the direction in which connectionbetween the liquid sucking/discharging line and the chip is maintained,and also the hook is energized by a lock releasing body provided at aspecified position in the direction in which locking between the liquidsucking/discharging line and the chip is released, so that it ispossible, when a filter holder is set in or removed from a tip sectionof a chip, to prevent the chip from being separated from the liquidsucking/discharging line without fail, and also locking with the hookcan automatically be released.

[0057] In the present invention, in a case of a liquid processingapparatus with the hook attached thereto, the filter holder set in a tipsection of the chip is transferred so that the chip and/or the filterholder is separated from an edge of the liquid sucking/discharging lineor a chip set therein when the liquid sucking/discharging line locked bythe locking body is moved upward, so that the work of removing a chipand/or a filter holder can be automated.

[0058] In the present invention, the vessel used according to thepresent invention is formed into a cassette form having a plurality ofchambers each for accommodating a type of liquid therein and thensamples or reagents required for a reaction or processing can bepipetted to each of the liquid accommodating section, so that highprecision liquid processing can be realized. In this case, a portion orall of the reagent previously accommodated in each liquid accommodatingsection is shielded with a thin film body which can be broken by theliquid sucking/discharging line or a chip set thereon, and a mechanismfor pipetting each reagent becomes unnecessary, which is desirable forsimplifying configuration of an apparatus.

[0059] In the present invention, in a case where the magnetic body isbuilt with a permanent magnet, a surface of the permanent magnetcontacting a chip is formed according to an external form of the chipand the chip is movably provided in a direction perpendicular to thelongitudinal direction of the chip, so that it is possible not only tocompletely capture magnetic particles, but also to prevent adverseeffects by diffusion and movement of the magnetic particles inassociation with the magnet without fail.

[0060] In the invention, the magnetic body is built with an electricmagnet in place of the permanent magnet described above with a surfacethereof contacting a chip formed according to an external form of thechip, and is provided so that the electric magnet generates a magneticforce when it contacts outside of the chip and also can move, whendegaussed, in a direction perpendicular to the longitudinal center lineof the chip or in a range including the direction, and for this reasonmagnetic particles are attracted in association with movement of themagnetic body along the longitudinal center line of the chip so that itis possible to prevent the magnetic particles from going out of controland control over the magnetic particles from being lost, which makes itpossible to realize complete attraction of the magnetic particles.

[0061] In the present invention, a holding body, which moves, when thepermanent magnet or electric magnet moves to the chip, in synchronism tomovement of the magnet, is provided, a surface of the holding bodycontacting a chip is formed according to an external form of the chip,and the chip is held between the holding body and the permanent magnetor electric magnet, so that the chip can easily be removed only bymoving the liquid sucking/discharging line upward.

[0062] In the present invention, a temperature control step required fora reaction between a target high molecular substance and a reagent orthe like or for amplifying the target high molecular substance isinserted into the liquid processing step with the liquidsucking/discharging line, the reaction liquid or the liquid to beamplified is transferred with the chip to each thermostatic vessel keptat a prespecified temperature, and also a covering body, which can beset in a tip section of the liquid sucking/discharging line, is set bythe liquid sucking/discharging line on each thermostatic vessel in whichthe reaction liquid or the liquid to be amplified is accommodated, sothat also amplification of the target high molecular substance cansuccessively be processed in a series of works.

[0063] Furthermore in the present invention, the covering body comprisesa flat surface section having a diameter larger than that of a bore ofthe thermostatic vessel and a maintenance groove section formed in asubstantially central portion of the flat surface section and having thesame bore as an external diameter of the liquid sucking/discharging lineor the tip of the chip, and a bottom section of the maintenance groovesection is formed with a thin film body which can be broken by theliquid sucking/discharging line or the chip, so that it is not necessaryto separately provide a covering body supply means or a liquidsucking/discharging means, which largely simplifies this type ofapparatus.

BRIEF DESCRIPTION OF DRAWINGS

[0064]FIG. 1 is a flat explanatory view showing general configuration ofan apparatus for extracting, recovering, and isolating DNA according tothe first embodiment of the present invention;

[0065]FIG. 2 is a cross-sectional view showing general configuration ofa nozzle unit in the apparatus;

[0066]FIG. 3 is an explanatory view explaining a step of removing a chipwith a hook body in the nozzle unit;

[0067]FIG. 4 is a flat explanatory view showing a state before U-shapedbody for removing the chip and the chip are engaged with each other;

[0068]FIG. 5 is a flat explanatory view showing a state after theU-shaped body for removing the chip and the chip are engaged with eachother;

[0069]FIG. 6 is a perspective view showing a case where the U-shapedbody is formed with a locking release rod;

[0070]FIG. 7 is a cross-sectional view showing an example of forms oftwo chips used in the present invention;

[0071]FIG. 8 is an explanatory view showing configuration of a pipettenozzle and a positional relation between a holding body and a magneticbody when getting closing to or away from each other;

[0072]FIG. 9 is a flat explanatory view showing configuration of amagnetic body as well as that of a holding body used in this embodiment;

[0073]FIG. 10 is an explanatory view showing positions at which themagnetic body and holding body are attached respectively;

[0074]FIG. 11 is a cross-sectional view for explanation of a relationbetween a bore of an intermediate diameter section of a chip and a widthdimension of a magnetic body;

[0075]FIG. 12 is a flow chart for explanation of steps from step 1 tostep 10 in the works of extracting, recovering, and isolating DNA withthe apparatus;

[0076]FIG. 13 is a flow chart for explanation of steps from step 12 tostep 19 in the works of extracting, recovering, and isolating DNA withthe apparatus;

[0077]FIG. 14 is a perspective view for explanation of an example ofconfiguration of a chip removing body;

[0078]FIG. 15 is a perspective view for explanation of another exampleof a means for engagement between a filter holder and a cell;

[0079]FIG. 16 is a cross-sectional view showing a case where a chip andtwo pieces of filter holder are connected and used in a multiple-stageform;

[0080]FIG. 17 is a cross-sectional view showing configuration of afilter holder with a filter having a large filter area attached thereto;

[0081]FIG. 18 is a cross-sectional view of a thermostatic vessel and acovering body used in this embodiment;

[0082]FIG. 19 is an explanatory view showing a state where a thin filmbody of the covering body is broken by a chip and a DNA amplifyingliquid accommodated therein is sucked;

[0083]FIG. 20 is a flat view for explanation of a state where a filterholder and a cell according to this embodiment are set in a cassette foreach group used in the reaction processing step;

[0084]FIG. 21 is a flat view for explanation of general configuration ofan apparatus for extracting, recovering, and isolating DNA comprising aplurality of reaction lines according to a second embodiment of thepresent invention;

[0085]FIG. 22 is a front view showing a quadrupled nozzle cylinderapplicable to the present invention,

[0086]FIG. 23 is a perspective view showing an example of configurationof a holding body and a magnetic body used in processing with thefour-stage nozzle cylinder, and

[0087]FIG. 24 is an explanatory view for operations of the holding bodyas well as of the magnetic body.

BEST MODE FOR CARRYING OUT INVENTION

[0088] Detailed description is made for the present invention withreference to embodiments thereof shown in the related drawings.

[0089]FIG. 1 shows an example of configuration in which the presentinvention is applied to an apparatus for extracting, recovering, andisolating DNA.

[0090] This apparatus comprises a pipette nozzle P supported by a nozzleunit so that it can freely be moved by an XYZ moving mechanism in thevertical direction as well as in the horizontal direction, chips T₁, T₂,T₃, and T₄ arranged from left to right in the FIG. 1, a chip removingbody E, a sample vessel C₀, a first filter holder H₁, a cell C₁, a cellC₂, a cell C₃, a second filter holder H₂, a cell C₄, a cell C₅, a cellC₆, a cell C₇, a thermostatic cell C_(8A), a cell C_(8B), a cell C_(8C)and a DNA recovery cell C₉.

[0091] Namely in this embodiment, the chips T₁, T₂, T₃ have a formsuited for holding the filter holders H₁, H₂, and the chip T₄ has a formsuited for capturing magnetic particles. It should be noted that,although the description of the embodiment above assumes a case where achip for capturing magnetic particles is processed with only one chipT₄, the form is not limited to that described above, and that aplurality of chips may be used according to the needs in the processingstep.

[0092] Also it should be noted that also in this embodiment, up to thefirst filter holder H₁, cell C₁, second filter holder H₂, cell C₂, cellC₃, cell C₄ controls the step of refining with a filter.

[0093] Also the cell C₅, cell C₆, and cell C₇ controls the magneticreaction as well as the steps of extracting and isolating, andfurthermore the thermostatic cell C_(8A), cell C_(8B), and cell C_(8C)control temperature in the reaction.

[0094] As described above, by arranging the cells sequentially,processing can be executed in a state where a plurality of specimens ofa sample are arranged in an array, and also control for driving thepipette nozzle P can be simplified. It is needless to say thatarrangement of the cells may be combined or changed according to theprocessing sequence.

[0095] It is desirable that the pipette nozzle P is connected directlyor with a certain but small range to a cylinder which can strictlycontrol a sucking/discharging rate with a servo motor or a pulse motorand unitized therewith.

[0096] A nozzle unit J supporting this pipette nozzle P comprises, asshown in FIG. 2, a vertical movement guide body 1 movably supported inthe XY direction (horizontal direction), a holder 2 connected with thevertical movement guide body 1 and moving in the vertical direction, asupporting body 3 extending in the horizontal direction from this holder2, the pipette nozzle P penetrating this supporting body 3 in thevertical direction and supported thereby, a spring 4 provided in thesupporting body 3 and energizing the pipette nozzle P in the downwarddirection in its normal state, and a hook bodies 6, 6 rotatablysupported on the opposite side to a lower projecting section 5 of thesupporting body 3. It should be noted that the sign Z in the figureindicates a sensor controlling a downward moving rate of the pipettenozzle P.

[0097] The hook bodies 6, 6 are energized in its normal state in theclosing direction due to an energizing force of the sheet springs 7, 7fixed to the lower projecting section 5. It should be noted that thespring 4 above is provided as a cushion for the pipette nozzle P, sothat it may be provided in any portion of the pipette nozzle P or thesupporting body 3, and also the sheet springs 7, 7 may directly beattached to the pipette nozzle P.

[0098] The nozzle unit having the configuration as described above isbuilt so that it can move in the XYZ directions (in the horizontaldirection and in the vertical direction) with the chips T₁, T_(2,) T₃engaged in a tip section of the pipette nozzle P supported by thisnozzle unit J, and this engagement is maintained because the hook bodies6, 6 locks a flange 8 Of the chips T₁, T₂, and T₃ in the state in whichthe hook body holds and embraces the chips.

[0099] The chip removing body E for releasing connection (locked state)with the hook bodies 6, 6 between the pipette nozzle P and the chips T₁,T₂, and T₃ comprises a pair of lock releasing rods 9, 9 provided at aposition for aborting chips as shown in FIG. 2 and FIG. 3, a U-shapedbody 10 having, for instance, a sheet form provided at the bottom of thelock releasing rods 9, 9 as shown in FIG. 4 and FIG. 5, and depositiontank (not shown) into which the chips T₁, T₂, T₃ separated from thepipette nozzle P are aborted.

[0100] As described above, the pipette nozzle P and chips T₁, T₂, T₃ arelocked and supported by the hook bodies 6, 6 to prepare against adischarging pressure of the liquid, or because the engagement betweenthe pipette nozzle P and the chips T₁, T₂, T₃ should not have beenreleased when separating the filter holders H₁, H₂ from the chips T₁,T₂, T₃.

[0101] For this reason, when releasing connection with the hook bodies6, 6 between the pipette nozzle P and the chips T₁, T₂, T₃, at first thenozzle unit J is moved downward at a position where the chip removingbody E is provided.

[0102] Then horizontal flange sections 6 a, 6 a of the hook bodies 6, 6contact the rock releasing rods 9, 9, and when the nozzle unit J movesfurther downward, the hook bodies 6, 6 rotate in the opening directionaround shafts 11, 11 as a support as shown in FIG. 3, and the lockedstate between the pipette nozzle P and the chips T₁, T₂, T₃ arereleased.

[0103] From the state described above, the nozzle unit J moves in thehorizontal direction, and as shown in FIG. 4, the flange 8 of each ofthe chips T₁, T₂, T₃ moves to a lower surface of the U-shaped body 10with a body section of the pipette nozzle P engaged in a U-shaped groovesection 12 of the U-shaped body 10, and then, when the nozzle unit Jgoes upward, the flange 8 of the chips T₁, T₂, T₃ contact a peripheralsection of the U-shaped groove section 12 of the U-shaped body 10, theupward movement thereof is restricted, so that only the pipette nozzle Pgoes upward and the chips T₁, T₂, T₃ are scraped away from the pipettenozzle P.

[0104] The description of the U-shaped body 10 above assumes a casewhere the U-shaped groove section 12 is provided in a plate body, butthe same effect can be achieved by connecting edge sections 13 of thelock releasing rods 9, 9 in a U-shaped form and forming them into amonolithic form.

[0105] This lock releasing rods 9, 9 and the U-shaped body 10 may beprovided not only at a position where the chip removing body E isprovided, but also at any of necessary positions described later.

[0106] By the way, in this embodiment, the chips T₁, T₂, T₃ are used asa set. It is needless to say that the number of chips may be increasedor reduced according to the processing sequence. Also a number of cellC₆ and cell C₇ is not limited to that in the embodiment shown in thefigure, and may be increased or reduced according to the necessity.

[0107] And formed in a lower edge section of the pipette nozzle P are 2stage sections P_(A), and P_(B) as shown in FIG. 7, and the chips T₁,T₂, T₃ are detachably set in the first stage section P while the chip T₄is detachably set in the second stage section P_(B).

[0108] Furthermore in the lower edge section of the pipette nozzle P, asshown in FIG. 8, a surface with flanges P_(C), P_(C) each projectingoutward from the peripheral surface and a surface P_(D) crossing theflanges P_(C), P_(C) are formed into a flat surface respectively.

[0109] As described above, by providing the flanges P_(C), P_(C) in aprojecting form, it is possible to remove the chips T₁, T₂, T₃ with theremoving body E, and also by having a surface P_(D) formed into a flatsurface form, it is possible to remove the chip T by holding it betweenthe holding body V and a magnetic body M.

[0110] In this holding body V and the magnetic body M, a known gearmechanism or a cum mechanism is, as show in FIG. 9, constructed so thatit opens or closes in synchronism to an operation of an opening/closingmechanism L comprising a rack and pinion mechanism or the like, and theopening/closing mechanism L is provided in a lower section of thecylinder unit S as shown in FIG. 10.

[0111] Also in the holding body V, a surface contacting the chip T₄ isformed into a concave form according to an external form of anintermediate diameter section K₁₂ of the chip T₄, and by holding thechip T₄between the holding body V and the magnetic body M and thenmoving the pipette nozzle P upward, it is possible to easily remove thechip T₄.

[0112] Setting of each of the chips T₁, T₂, T₃, and T₄ having theconfiguration as described above is executed, for instance, bytransferring the pipette nozzle P to just above a chip rack (not shown)with the chips T₁, T₂, T₃, T₄ supported thereon in at an erectedposition, then descending the pipette nozzle P, and press-fitting thelower edge section PA or PB of the pipette nozzle O into upper edgesections of the chips T₁, T₂, T₃, T₄.

[0113] Namely each of the chips T₁, T₂, T₃ each with the filter holdersH₁, H₂ set thereon comprises, as shown in FIG. 7, a small diametersection K₁ an intermediate diameter section K₂ connected to an uppersection of this small diameter section K₁, and a large diameter sectionK₃ connected to an upper section of this intermediate diameter sectionK₂, all of which are connected to each other in the vertical directionand formed into a monolithic form, and the filter holder H₁, H₂described later are detachably set in the intermediate diameter sectionK₂.

[0114] The intermediate diameter section K₂ of each of the chips T₁, T₂,T₃has the substantially same diameter as or a slightly larger diameterthan an inner diameter of the engaging section between the filterholders H₁, H₂, and length of the small diameter section K₁ is shortenough for tip sections of the filter holders H₁, H₂ not to contact afilter when the filter holders H₁, H₂ are engaged.

[0115] On the other hand, the chip T₄to which magnetic particles areattracted is different in the purpose of its use as well as in themethod of using it from the chips T₁, T₂, T₃ with filter holders setthereon respectively, and the chip T₄ comprises, as shown in FIG. 7, thelarge diameter section having an internal bore engaged onto the secondstage section P_(B) which is smaller than the first stage section P_(A)of the pipette nozzle P, the intermediate diameter section K₁₂ having adiameter smaller than that of the large diameter section K₁₃, and thesmall diameter section K₁₁ having a diameter smaller than that of theintermediate diameter section K₁₂, and a magnetic body M attractingmagnetic particles is moved to or away from the intermediate sectionK₁₂.

[0116] An internal diameter of the intermediate diameter section K₁₂ ofthis chip T₄ has a dimension enough for a ferromagnetic field of themagnetic body M to be effective therein, and preferably the dimensionis, as shown in FIG. 11, substantially identical to a width dimension ofthe surface of the chip to which the magnetic body M contacts.

[0117] It should be noted that, although the description of the presentembodiment above assumes a case where, from each of the chips T₁, T₂, T₃only the filter holders are removed, but in a case of a reaction step inwhich only the filter holders H₁, H₂ are not required to be removed,locking with a hook body is not required, so that a bore of the largediameter section of a chip for a filter holder may be the same as thatof the chip T₄.

[0118] The apparatus for extracting, recovering, and isolating DNAhaving the configuration as described above is driven and control in thesteps shown in FIG. 12 and FIG. 13.

[0119] At first, as shown in FIG. 12, in step 1, the first chip T₁ isinserted into the first stage P_(A) in a lower edge section of thepipette nozzle P. In this step, the pipette nozzle P and the chip T₁ arelocked by the hook bodies 6, 6 and the connection between the twomembers are maintained.

[0120] When the chip T₁ has been set in the first stage P_(A) in thelower edge section of the pipette nozzle P, the pipette nozzle P istransferred to just above the sample vessel C₀ with a sampleaccommodated therein and then goes down with the liquid surface checkedby a liquid surface sensor Z₁ (Refer to FIG. 10), and then the loweredge section of the chip T₁ is inserted into the sample and a requiredquantity of the sample is sucked (step 2).

[0121] The sample used in this embodiment is natural blood having beensubjected, before start of experiment, to cell core lysis or proteinlysis with such a solution as an SDS solution or a proteinase Ksolution, but a step of cell core lysis or protein lysis using the abovesolution may be incorporated in this processing flow.

[0122] Then the chip T₁ having absorbed a required quantity of sampletherein is transferred to just above the first filter holder H₁ with thefirst filter F₁ provided therein, and goes down with the first filterholder H₁ locked and engaged in a lower edge section of the chip T₁(step 3).

[0123] The filter F₁ set in this first filter holder H₁can remove bloodcorpuscle shells in the sample above from the blood dissolved anddischarge a lymphocyte solution containing DNA into the cell C₁. Thepipette nozzle P with the first filter holder H₁ locked and engaged inthe lower edge section of the chip T₁ is then transferred to just abovethe cell C₁, and at the position the pipette nozzle P starts adischarging operation, separates cell membrane and blood corpuscleshells in the sample from lymphocytes and DNA loading reburied pressureto the sample sucked into the chip T₂, and discharges only thelymphocytes and DNA into the cell C₁ (step 4). At this point of time, asthe pipette nozzle P is energized downward by the spring 4, the flange13 of the first filter holder H₁ is pressed and closely adhered to aperiphery of an opening of the cell C_(1,) so that generation of leakdue to a discharging pressure of the liquid is prevented.

[0124] Then the pipette nozzle P is transferred to just above a positionwhere the chip removing body E shown in FIG. 1 is provided, the chip T₁and first filter holder H₁ are removed from the lower edge section ofthe pipette nozzle P according to the processing sequence describedabove (step 5), and the chip T₁ and first filter holder H₁ removed asdescribed above are aborted into a waste tank (not shown).

[0125] Then the pipette nozzle P is transferred to just above a chiprack with the chip supported in the erected position, and goes down atthe position with the second chip T₂ set in the first stage P_(A) in thelower edge section of the pipette nozzle P (step 6). Also in this case,the pipette nozzle P and the chip T₂ are locked and connected to eachother by the hook bodies 6, 6.

[0126] Then the pipette nozzle P with the chip T₂ set therein istransferred to just above the cell C₁ again and goes down at theposition with a required quantity of lymphocyte solution sucked from thecell C₁ there (step 7).

[0127] Then the pipette nozzle P is transferred to just above the secondfilter holder H₂ with the second silica membrane filter F₂ providedtherein and goes down there with the second filter holder H₂ locked andengaged in a lower edge section of the chip T₂ (step 8).

[0128] Then the silica membrane filter F2 set in this second filterholder H₂ separates DNA from foreign materials in the lymphocytesolution above, and discharges the residual solution into the cell C₂.

[0129] As described above, the pipette nozzle P with the second filterholder H₂ locked and engaged in the lower edge section of the chip T₂ istransferred to just above the cell C₂, and at the position the pipettenozzle P starts an discharging operation, separates DNA from foreignmaterials in the lymphocyte solution loading required pressure to thelymphocyte solution sucked into the chip T₂, and discharges the residuallymphocyte solution into the cell C₂ (step 9). In this step, as thepipette nozzle P is energized downward by the spring 4, the flange 14 ofthe second filter holder H₂ is pressed and closely adhered to aperiphery of an opening of the cell C₂, so that generation of leak dueto a discharging pressure of the liquid is prevented.

[0130] Then the pipette nozzle P with the second filter holder H2 havingcaptured DNA and locked and engaged therein is transferred to just abovethe third cell C₃ in which a cleaning liquid is accommodated, and goesdown there with the second filter holder H₂ steeped into the cleaningliquid in the cell C₃ (step 10).

[0131] Then the locked state of the chip T₂ and filter holder H₂ isreleased by the filter holder removing body E₁ shown in FIG. 14, andonly the second filter holder H₂ is steeped into the cleaning liquid inthe cell C₃ (step 11). It should be noted that the chip T₂ istransferred to just above the chip removing body E, removed from thepipette nozzle P, and is aborted according to the sequence describedabove.

[0132] It should be noted that the filter holder removing body E₁ has aflat surface with a notched groove 15 having a substantially U-shapedform, and a diameter of the notched groove 15 is slightly larger than anexternal diameter of a body section of each of the chips T₁, T₂, T₃, butis smaller than a diameter of the flanges 13, 14 of the filter holdersH₁, H₂.

[0133] Then the pipette nozzle P is transferred, as shown in FIG. 13, tojust above a chip rack with the chip supported in the erected positiontherein, and at the position goes downward with a third chip T₃ set inthe first stage PA in the lower edge section of the pipette nozzle P(step 12). Also in this step, the pipette nozzle P and chip T₃ arelocked by the hook bodies 6, 6 and the connection is maintained.

[0134] Then the pipette nozzle P with this chip T₃ set therein is gaintransferred to just above the cell C₃, and goes downward at the positionwith the second filter holder H₂ locked and engaged in a lower edgesection of the chip T₃ (step 13).

[0135] Then the pipette nozzle P starts a sucking operation, and arequired quantity of a mixture liquid of a cleaning liquid and DNA issucked. With this operation, the work of refining DNA is complete.

[0136] It should be noted that, in the present embodiment, as a meansfor steeping the second filter holder H₂ into a cleaning liquidaccommodated in the cell C₃, for instance a locking projection W, whichallows intrusion of the second filter holder H₂ but prevents it fromgoing off may be provided in the cell C₃ in place of the removing body Eas shown in FIG. 15 with also a locking projection S engaging with thelocking projection W provided for the purpose to steep the second filterholder H₂ into a cleaning liquid accommodated in the cell C₃ by havingthe locking projection W and locking projection S engaged with eachother.

[0137] Description of the embodiment shown in the figures assumes a casewhere a number of filter holders set in each of the chips T₁, T₂, T₃ isonly one, but in the present invention the filter holder H₁ (or H₂) maybe set in each of the chips T₁, T₂, T₃ with the filter holder H₂ (or H₁)set in the filter holder H₁ (or H₂) to form a two-stage joint of filterholders, and also that a number of stages may be two or more accordingto the necessity in the processing sequence.

[0138] Furthermore, in certain types of liquid processing, a filter areamay be short in the embodiment as shown in the figures, and in thiscase, as shown in FIG. 17, a filter accommodating section Q having adiameter larger than that of the body section and also having anexpanding and projecting form may be provided in the intermediatesection of the filter holder H₃ to accommodate the filter F₃ having alarge filter area in the filter accommodating section Q. An externaldiameter of the filter accommodating section Q in this case shouldpreferably be smaller than a diameter of a projections C_(B), C_(B) forpositioning projecting from the upper edge flange section C_(A) of thecell C. It should be noted that the sign R in the figure indicates astay member for supporting the filter F₃ in an intermediate section ofthe filter accommodating section Q. It is needless to say that thefilter F3 may be supported by mesh in its lower section.

[0139] Next description is made for a step of subjecting DNA refinedthrough the reaction steps described above to such works as extracting,recovering, isolating or amplifying with PCR or to control fortemperature thereof.

[0140] Namely, in a case where such works—as extracting, recovering, orisolating by making use of this pipette device with magnetic particles Gwith DNA or DNA-bonded substance bonded to the surface, as shown in step14 in FIG. 13, at first the pipette nozzle P is moved upward and thentransferred to just above a fourth cell C₄ with the second filter holderH₂ left in cell₃ via a filter holder removing body E₂ having the sameconfiguration as that of the filter holder removing body E₁ and thesucked DNA solution is discharged into the cell C₄.

[0141] A required quantity of reaction liquid containing magneticparticles G with DNA or DNA-bonded substance bonded to the surfacethereof has been supplied into this cell C₄, and when the DNA solutionis discharged into the reaction liquid, a reaction between DNA fragmentsand the magnetic particles G is started.

[0142] The chip T₃ with the DNA solution having been discharged into thecell C₄ is removed from the lower edge section of the pipette nozzle Paccording to the processing sequence like in a case of the chip T₁ orchip T₂, and is aborted.

[0143] It is needless to say that then the chip T₄ is set in the loweredge section of the pipette nozzle P according to the processingsequence as described above.

[0144] Then, after a certain period of time has passed, the pipettenozzle P goes downward and steeps the chip T₄ into the reaction liquid,the magnetic body M contacts the intermediate diameter section K₁₂ ofthe chip T₄, the works of sucking and discharging the liquid by thepipette nozzle P is executed at least once according to the necessity,and separation between the magnetic particles and the reaction liquid isexecuted (step 15). Then the sucking and discharging work is executed toa slow speed so that almost all the magnetic particles are captured. Inthis case, it is important for completely attracting the magneticparticles to provide controls over the sucking and dischargingoperations so that the final liquid surface of the reaction liquidsucked or discharged passes through an area effected by a magnetic forcegenerated by the magnetic body M.

[0145] With this separating operation, only the magnetic particles Gwith DNA bonded thereto are attracted almost completely onto an internalsurface of the chip T₄, and the residual liquid is discharged into thecell C₄.

[0146] In this embodiment, the processing is executed by moving themagnetic body M upward and backward simultaneously in a directionperpendicular to the longitudinal direction of the chip T₄ or turningON/OFF an electric magnet.

[0147] In a case where processing is executed with an electric magnet,the electric magnet is controlled so that the electric magnet is turnedON to generate a magnetic force magnet when it contacts an externalsurface of the chip T₄ and is moved upward and backward simultaneously,when degaussed, in a direction perpendicular to the longitudinaldirection of the chip T₄.

[0148] Then this chip T₄ with magnetic particles G attracted onto theinternal surface is transferred to a cell T₅ in which such a reagent asrestriction enzyme liquid or the like required for extracting,recovering, and isolating the target DNA, and at the position operationsfor sucking or discharging the reagent such as the restriction enzymeliquid are executed by pumping as described above (step 16). Theoperations for sucking and discharging the reagent are executed severaltimes continuously with a tip section of the chip T₄ steeped into thereagent, so that intrusion of bubbles is prevented. In this step, bysetting the magnetic body M so that a magnetic force generated by themagnetic body M is not effective, works of mixing and agitating thereagent such as a restriction enzyme liquid and the magnetic particlescan be executed with high precision, and an excellent reaction state canbe insured.

[0149] After the reagent such as a restriction enzyme liquid andmagnetic particles are fully agitated and mixed with each other, thechip T₄ again sucks and discharges this liquid slowly, and executes theoperations once or required times according to the necessity, andseparation of magnetic particles from the liquid with the magnetic bodyM is executed.

[0150] With the operations, only the magnetic particles G with DNAbonded thereto are almost completely attracted onto an internal surfaceof the chip T₄, and the residual liquid is discharged into the cell C₅(step 17).

[0151] Then the chip T₄ with magnetic particles G attracted onto theinternal surface thereof is transferred to cells C₆, C₇ by turns, inwhich a reagent required for extracting, recovering, and isolating atarget DNA is previously accommodated, and at the positions where thecells C₆, C₇ are provided, and the reaction processing by pumping isexecuted like in the cases described above (step 18).

[0152] In this step, by setting the magnetic body M in the state whereina magnetic force generated by the magnetic body M is not effective,works of agitating and mixing the reagent and magnetic particles G witheach other can be executed with high precision. It is needless to saythat times of pumping is not limited to the values employed in the abovecases, and can be increased or reduced according to the necessity.

[0153] Also in a case where temperature control or amplification isrequired during the processing described above, if it is required tomaintain the temperature at 90° C., 60° C., or 40° C., the reactionliquid may be transferred to the thermostatic cells C_(8A), C_(8B),C_(8C) each heated to the target temperature. In this case, as comparedto a case where temperature control is executed with one unit of heatingmeans like in the conventional technology or a case where a solution istransferred vessel by vessel to the heating section, the reaction can beexecuted effectively, and also amplification under controlledtemperature can easily be executed within a short period of time, andfurther a device for transferring vessels is not required, so that theapparatus can be simplified.

[0154] Furthermore, if a sample is to be heated to a high temperaturesuch as 60° C. or 90° C., a mixture solution is evaporated, and toprevent the evaporation, in this embodiment, as shown in FIG. 18, it isdesirable to set a covering body L.

[0155] This covering body L is engaged and locked in a thermostatic cellC₈ to be heated to a high temperature of the thermostatic cells C_(8A),C_(8B), C_(8C) accommodated in a vessel accommodating hole provided in aheating member such as a heater block, and comprises a flat surfacesection L₁ having a diameter larger than a bore of the thermostatic cellC₈, a locking piece section L₂ having a substantially L-shaped form andengaging with a locking projection Y₁ extending downward from aperiphery of this flat surface section L₁ and projecting to above anexternal periphery of the thermostatic cell C₈, a supporting groove L₃provided in a concave form at a center of the flat surface section L₁, atin film section L₄ closing a bottom section of the supporting groovesection L₃ and formed with aluminum or a similar material, and a sealprojecting section L₅ projecting from an external periphery of thesupporting groove section L₃, and the supporting groove section L has abore which is identical to an outer diameter of a tip of the pipettenozzle P.

[0156] It should be noted that the thin film section L₄ described abovemay be formed by heating and welding a separate seal material such asaluminum to the supporting groove section L₃ or by means of supersonicwelding, or may be formed with soft plastics, which is the same materialas that for the supporting groove section L₃, into a thin film form.

[0157] For the reasons as described above, after a mixture solution ispoured into the thermostatic cell C₈, the pipette nozzle P with the chipT₄ having been removed therefrom is transferred to a position where thecovering body L is stocked, then goes down with the tip section of thepipette nozzle P press-fit into the supporting groove section L₃ of thecovering body L, and the pipette nozzle P is transferred with thecovering body L supported thereon to just above the thermostatic cellC₈, where the locking piece section L₂ of the covering body L is engagedwith a locking projection Y₁ of the thermostatic cell C₈. It is needlessto say that the thermostatic cell C₈ is locked so that it is not raisedfrom a heating member such as a heater block.

[0158] After the work as described above is finished, the pipette nozzleP goes upward, and in this step, the covering body L is fixed to thethermostatic cell C₈ so that it is not removed from the thermostaticcell C₈, and for this reason the tip section of the pipette nozzle Pgoes out of the supporting groove section L₃ in the covering body L, andonly the pipette nozzle P is moved to a prespecified position.

[0159] Then a new chip (not shown) is set to the tip section of thepipette nozzle P, and the pipette nozzle P is again transferred to justabove the thermostatic cell C and goes down there with the tip sectionof the chip T inserted into the supporting groove section L₃ of thecovering body L and breaking through the thin film section L₄ downward,and then the pipette nozzle P sucks a mixture solution accommodated inthe thermostatic cell C₈ thereinto, goes upward, and sends the suckedmixture solution to the next thermostatic cell C₈ or cell C₉.

[0160] Thus, all of the DNA solution sucked into the cell C₇ or thethermostatic cell C₈A is discharged into the cell C₉. Then the magneticbody M contacts the intermediate diameter section K₁₂ of the chip T₄,operations for sucking and discharging by the pipette nozzle P areexecuted once or required times to separate the magnetic particles Gfrom the DNA solution, and only the DNA solution is discharged with themagnetic particles G attracted onto an internal surface of the chip T₄(step 19).

[0161] The above description of the first embodiment of the presentinvention assumes a case where the filter holders H₁, H₂, sample cellC₀, and cells C₁ to cell C₉ are arrayed in the order of reaction steps,but the present invention is not limited to this configuration, and asshown in FIG. 20, excluding the sample cell C₀ and DNA recovery cell C₉each of a group of cells C₁ to C₄ used for refining with a filter, agroup of filter holders H₁, H₂, a group of cells C to C₇ processed withthe magnetic particles G, and a group of thermostatic cells C_(8A) toC_(8C) may be set in a cassette with the pipette nozzle P driven andcontrolled according to the processing sequence described above. It isneedless to say that the covering body L may be provided in parallel toa cassette of the thermostatic cells C_(8A) to C_(8C).

[0162]FIG. 21 shows a second embodiment of the present invention, andthis embodiment shows a case where a plurality of reaction lines, forinstance, 4 arrays of reaction lines each having the same configurationas that of the single reaction line are provided and the lines areseparated from each other with a partition wall X. It is needless to satthat, in this case, a required number of pipette nozzles are provided inseries for each reaction line so that a plurality of samples cansimultaneously be processed.

[0163] Also it should be noted that the reagent vessels Ra, Rb, Rc, Rd,Re, Rf arrayed along each of the reaction lines above and chips T_(5A),T_(5B), T_(5C), T_(5E), T_(5F) each for pipetting each reagent areprovided in parallel along the trajectories of the pipette nozzles P₁,P₂, P₃, P₄ moving along each of the reaction lines.

[0164] The partition wall X comprises a rectangular plate body having asize in a range including a tip section of each of the pipette nozzleP₁, P₂, P₃, P₄ when pulled up, and this partition wall X forms a workingspace separated from a neighboring reaction line, and with thisconstruction it is possible to prevent foreign materials other thantarget DNA from coming in from other reaction lines.

[0165] It should be noted that an air inhalator (not shown) having along air inhalating port in the longitudinal direction of each reactionline may be provided between each reaction line for air inhalation inplace of the partition wall X.

[0166] With this construction, a curtain of a downward air flow isgenerated in each line, and the same working space as that formed withthe partition wall X described above is formed, so that it is possibleto prevent foreign materials other than the target DNA from coming infrom other reaction lines without fail. In a case where an air inhalatoris used, there exists no physical curtain, so that a form and movementof a pipette nozzle allowable in the production line is relatively free.Furthermore the air inhalator may be provided above each reaction line,and in this case a curtain of upward air flow is generated between thereaction lines, and a similar working space is generated between eachline. It is needless to say that this air inhalating method may be usein combination with the partition wall X described above, and in thatcase cross contamination between the reaction lines can be preventedwith higher accuracy.

[0167]FIG. 22 shows a cylinder J₁ used in the second embodiment of thepresent invention, and this cylinder J₁ shows the configuration in acase where works in each reaction line are not executed with separatecylinders, but executed only with this cylinder J₁, and theconfiguration and effects thereof are similar to those provided by thistype of cylinder known in the conventional technology excluding thepoint that four chips T₁, T₂, T₃, T₄ or T_(5A), T_(5B), T_(5C), T_(5E)or the like used in the second embodiment above each detachably set inthe pipette nozzles P₁, P₂, P₃, P₄ respectively can simultaneously beset, and detailed description thereof is omitted herein. A number ofchips used in the present invention is not limited to four, and aplurality of chips may be set according to a number of liquid processinglines.

[0168]FIG. 23 and FIG. 24 show a mechanism suited to driving andcontrolling a magnetic body M and a holding body V when a liquid isprocessed with the cylinder shown in FIG. 22, and in this instance amagnetic body M having magnetic sections M₁, M₂, M₃, M₄ formed into ashape like comb teeth and a holding body V having holding sections V₁,V₂, V₃, V₄ also formed into a shape like comb teeth are pivotallysupported by an up-down mechanism O in a state where opening or closingthereof is freely executed, and this up-down mechanism O is moved upwardand downward, when rollers O_(R), O_(R) of the up-down mechanism O areclosed as shown in FIG. 24. The magnetic body M and holding body V aremoved and closed by the spring O_(S) shown in FIG. 23 to hold a chipbetween them, and as a result the magnetic body M is simultaneouslycontacted to the four chips T_(A), T_(B), T_(C), and T_(D), or each chipis simultaneously held by the holding body V and the magnetic body M.

[0169] The embodiment shown in FIG. 23 and FIG. 24 shows a case where aplurality of liquid processing lines are formed with partition lineslike in the second embodiment by providing the magnet body M and theholding body V, and in this case the magnetic body M and holding body Vnever collide against the partition wall, and such works as attracting,agitating, and mixing magnetic particles or sucking and discharging aliquid can simultaneously be executed in four liquid processing lines atthe same timing, so that the processing efficiency can substantially beimproved with simple configuration. It is needless to say that thepresent invention is not limited to a case where four pieces of magneticbody M and four holding bodies V are used like in the embodimentdescribed above, and that any number of magnetic bodied and holdingbodies may be used according to the necessity.

[0170] Also it should be noted that a filter may be set above a largediameter section of each chip to prevent a liquid from being depositedon or attracted to the sucking/discharging line.

INDUSTRIAL APPLICABILITY

[0171] As described above, the liquid processing method and theapparatus for the same making use of the pipette device according to thepresent invention is suited for executing such works as quantifying,separating, taking out, pipetting, cleaning, condensing, and diluting aliquid or a target high molecular substance contained in a liquid aswell as such works as extracting, recovering, and isolating the targethigh molecular substance automatically and with high precision by meansof sucking and discharging the liquid with a liquid sucking/dischargingline in the pipette device. The liquid processing method and theapparatus for the same described above is suited for, for instance,executing such works as quantifying, separating, taking out, pipetting,cleaning, condensing, diluting a useful substance such as antibiotics, agenetic substance such as DNA or an immunological substance such asantibodies, and/or such works as extracting, recovering, and isolating atarget high molecular substance automatically and with high precision bymeans of sucking and discharging the liquid in the liquidsucking/discharging line in the pipette device.

1. A liquid processing method making use of a pipette device which sucksa liquid containing a target high molecular substance via a chipdetachably set in a sucking port or a discharging port of a liquidsucking/discharging line from inside of a vessel and transfers theliquid or the target high molecular substance to a target nextprocessing position, wherein said chip has the sucked target highmolecular substance deposited on magnetic particles and/or separatedwith a filter set in the chip.
 2. A liquid processing method making useof a pipette device according to claim 1, wherein said target highmolecular substance is a useful substance such as antibiotics, a geneticsubstance such as DNA or an immunological substance such as antibody. 3.A liquid processing method making use of a pipette device according toany of claim 1 or claim 2, wherein such works as quantifying,separating, taking out, pipetting, cleaning, condensing, and dilutingsaid target high molecular substance is carried out with a chip set insaid liquid sucking/discharging line and at least one type of filter setin said chip.
 4. A liquid processing method making use of a pipettedevice according to claim 3, wherein a plurality of filter holders forholding a filter can be provided in multiple stages in said chip.
 5. Aliquid processing method making use of a pipette device according to anyof claim 3 or claim 4, wherein a plurality of filters held by saidfilter holders comprise one or a plurality types of filters each havinga different pore size for separation of a target high molecular materialand foreign materials other than the target high molecular substancerespectively.
 6. A liquid processing method making use of a pipettedevice, wherein after such works as quantifying, separating. taking out,pipetting, cleaning, condensing, diluting a liquid or a target highmolecular substance with the filter according to any of claim 3 to claim5, in a step of detachably setting a new chip in a tip section of saidliquid sucking/discharging line and sucking/discharging a solutioncontaining magnetic particles with this chip, said magnetic particlesare attracted by a magnetic body provided in the side of the chip ontoan internal surface of the chip to extract, recover, and isolate thetarget high molecular substance.
 7. A liquid processing method makinguse of a pipette device according to any of claim 1 or claim 2, whereinsuch works as capturing, extracting, isolating, amplifying, labelling,and measuring said target high molecular substance may be executed onlywith a chip set in said liquid sucking/discharging line, a magneticforce, and one or a plurality types of magnetic particles.
 8. A liquidprocessing method making use of a pipette device according to claim 7,wherein by causing the chip set in said liquid sucking/discharging lineto react with magnetic particles, refining process such as capturingcells, having cell cores or protein lysed is executed, and a particulartarget high molecular substance is extracted.
 9. A liquid processingmethod making use of a pipette device according to claim 7, wherein byusing magnetic particles with a probe or biotin or streptoavidin coatedthereon by making use of a chip set in said liquid sucking/dischargingline, a particular base sequence piece is isolated.
 10. A liquidprocessing method making use of a pipette device according to claim 7,wherein by causing the chip set in said liquid sucking/discharging lineto react with magnetic particles, refining process such as capturingcells, having cell cores or protein lysed is executed, and a particulartarget high molecular substance is extracted, then the particular basesequence piece is isolated with other type of magnetic particles with aprobe or biotin or streptoavidin coated thereon.
 11. A liquid processingmethod making use of a pipette device according to any of claim 7 toclaim 10, wherein after a series of works such as capturing, extracting,and isolating a target high molecular substance by using said magneticparticles, by making the isolated particular base sequence piece emitlight through chemical luminescence or fluorescence or enzymaticcoloration, presence or a quantity of the particular base sequence pieceis detected or measured.
 12. A liquid processing method making use of apipette device according to any of claim 7 to claim 10, wherein bycausing the chip set in said liquid sucking/discharging line to reactwith magnetic particles, refining process such as capturing cells,having cell cores or protein lysed is executed, and a particular targethigh molecular substance is extracted, then the extracted target highmolecular substance is amplified, the particular base sequence piece isisolated with other type of magnetic particles with a probe or biotin orstreptoavidin coated thereon and then presence of or a quantity of theparticular base sequence piece is detected and measured by causing theisolated piece to emit light through chemical luminescence orfluorescence or enzymatic coloration.
 13. A liquid processing methodmaking use of a pipette device according to any of claim 1 to claim 12,wherein the works of separating, taking out, pipetting, cleaning,condensing, diluting said target high molecular substance and/or worksfor capturing, extracting, isolating, amplifying, labelling, andmeasuring the substance are executed in a single liquidsucking/discharging line.
 14. A liquid processing method making use of apipette device according to any of claim 1 to claim 12, wherein theworks of separating, taking out, pipetting, cleaning, condensing,diluting said target high molecular substance and/or works forcapturing, extracting, isolating, amplifying, labelling, and measuringthe substance are executed in a plurality of liquid sucking/discharginglines provided in parallel to each other.
 15. A liquid processing methodmaking use of a pipette device according to claim 14, wherein saidplurality of liquid sucking/discharging lines execute the works ofseparating, taking out, pipetting, cleaning, condensing, diluting saidtarget high molecular substance and/or the works of capturing,extracting, isolating, amplifying, labelling, and measuring thesubstance in each line according to the same timing.
 16. A liquidprocessing method making use of a pipette device according to claim 14,wherein said plurality of liquid sucking/discharging lines execute theoperations of sucking, discharging each liquid by a different timingaccording to a processing step specified by each liquid or independenttiming.
 17. A liquid processing method making use of a pipette deviceaccording to any of claim 13 to claim 16, wherein working spaces in thesingle or a plurality of liquid sucking/discharging lines are separatedfrom each other with partitions.
 18. A liquid processing method makinguse of a pipette device according to any of claim 13 to claim 16,wherein air sucking ports each are provided in each line working spacerespectively in said single or a plurality of liquid sucking/discharginglines, and the working spaces are separated from each other with an airflow.
 19. A liquid processing method making use of a pipette deviceaccording to any of claim 13 to claim 16, wherein working spaces in saidsingle or a plurality of liquid sucking/discharging lines are separatedfrom each other with partitions, and air in the working spaces separatedfrom each other with the partitions is sucked from an air sucking portprovided in the working space.
 20. A liquid processing method making useof a pipette device according to any of claim 1 to claim 19, whereinsaid magnetic particles have a target high molecular substance or asubstance bonded to a target high molecular substance deposited orabsorbed to a surface of each magnetic particles.
 21. A liquidprocessing method making use of a pipette device according to any ofclaim 1 to claim 19, wherein said magnetic particles are absorbed ontoan internal wall of a chip due to a magnetic force working from outsideof said chip, and, if effect of said magnetic force is weak or notpresent, the magnetic particles can be separated from the internalsurface of the chip.
 22. A liquid processing method making use of apipette device according to any of claim 1 to claim 21, whereincontrolling load of a magnetic force into or elimination of a magneticforce in said chip is executed by means of moving a permanent magnet ina direction perpendicular to the longitudinal direction of the chip orin a range including the direction perpendicular to the longitudinaldirection of the chip.
 23. A liquid processing method making use of apipette device according to any of claim 1 to claim 21, whereincontrolling load of a magnetic force into or elimination of a magneticforce in said chip is executed by turning ON or OFF an electric magnet.24. A liquid processing method making use of a pipette device accordingto claim 23, wherein said electric magnet is driven and controlled sothat the electric magnet generates a magnetic force when it contacts anexternal surface of the chip and moves in a direction perpendicular tothe longitudinal direction of the chip or in a range including thedirection perpendicular to the longitudinal direction of the chip whenthe magnetic force is eliminated.
 25. A liquid processing method makinguse of a pipette device according to any of claim 21 to claim 24,wherein a holding body synchronously moves when the permanent magnet orelectric magnet moves to the chip, and said chip is held with saidpermanent magnet or electric magnet and holding body.
 26. A liquidprocessing method making use of a pipette device according to any ofclaim 1 or claim 6 to claim 25, wherein said chip comprises a smalldiameter section steeped into a liquid, a large diameter section havinga capacity larger than a capacity of a vessel in which a liquid isaccommodated, and an intermediate section provided between the smalldiameter section and the large diameter section and having a diametersmaller at least than that of the large diameter section, and magneticparticles are captured by said intermediate section.
 27. A liquidprocessing method making use of a pipette device according to claim 26,wherein an internal diameter of the intermediate section of said chiphas a dimension appropriate for ferromagnetic field of said magneticbody to provide effects therein, and magnetic particles are captured dueto a magnetic force generated in the ferromagnetic field of the magneticbody within a short period of time.
 28. A liquid processing methodmaking use of a pipette device according to claim 26 or claim 27,wherein an internal diameter of the intermediate section of said chip isformed so as to have the substantially same width dimension as that of acontacting surface of a magnetic body contacting said intermediatesection.
 29. A liquid processing method making use of a pipette deviceaccording to any of claim 1 or claim 6 to claim 28, wherein sucking ordischarging a liquid is controlled so that absorption of magneticparticles onto an internal surface of a chip set in said liquidsucking/discharging line is executed by passing a solution containingmagnetic particles through a ferromagnetic field inside a chip at a slowspeed appropriate for the magnetic particles to be captured completely.30. A liquid processing method making use of a pipette device accordingto claim 29, wherein controls are provided so that the final liquidsurface of a liquid passing through said chip when sucked into ordischarged from the chip always reaches said magnetic field.
 31. Aliquid processing method making use of a pipette device according to anyof claim 1 or claim 6 to claim 30, wherein, when sucking magneticparticles into said chip, a tip section of a chip set in the liquidsucking/discharging line is driven and controlled so that the chipsection contacts an internal bottom of a vessel with the liquidaccommodated therein and then lightly rises so as to suck the liquid.32. A liquid processing method making use of a pipette device, whereinagitation and mixing of magnetic particles absorbed in the said chipwith a reagent or cleaning water is achieved under conditions in whichthe works of sucking and discharging the liquid is continuously executedin the liquid sucking/discharging line at a high speed and times enoughto agitate and mix the liquid with the magnetic particles.
 33. A liquidprocessing method making use of a pipette device according to claim 32,wherein, when magnetic particles absorbed in the said chip are agitatedand mixed with a reagent or cleaning water, the works of sucking anddischarging the liquid in said liquid sucking/discharging line is drivenand controlled so that no bubble is generated in a state in which thetip section of the chip is steeped into a reagent or cleaning wateraccommodated in a vessel.
 34. A liquid processing method making use of apipette device, wherein controlling a temperature required for reactionbetween said target high molecular substance and reagent or the like oramplification of the target high molecular substance is executed bytransferring the reaction liquid or a liquid to be amplified into eachthermostatic vessel previously kept at a constant temperature with saidchip.
 35. A liquid processing method making use of a pipette deviceaccording to claim 34, wherein, when controlling a temperature in eachreaction vessel, a covering body is set over a tip section of saidliquid sucking/discharging line, and said covering body is set via theliquid sucking/discharging line on the thermostatic vessel.
 36. A liquidprocessing method making use of a pipette device according to claim 35,wherein said liquid sucking/discharging line or a chip set in said lineis driven and controlled so that said covering body is plucked andbroken and a reaction liquid or a liquid to be amplified in athermostatic vessel is sucked thereby.
 37. A liquid processing apparatusmaking use of a pipette device comprising a liquid sucking/dischargingline which can move in the horizontal direction and is maintained at aspecified position so that it can move in the vertical direction; ameans for executing liquid sucking/discharging works through said liquidsucking/discharging line; a plurality of chips required for processingone type of liquid and provided along the horizontal direction in whichthe liquid sucking/discharging line moves; a vessel with said liquidaccommodated therein; one or more filter holders each having a filterrequired for the processing above; and one or more vessels with othertypes of liquid accommodated therein respectively required for theprocessing above, and the liquid sucking/discharging line or a chip settherein is driven and controlled according to instructions from acontrol unit so that the line or the chip is transferred with a filterholder set therein to execute such works as quantifying, separating,taking out, pipetting, clearing, condensing, and diluting the liquid ora target high molecular substance contained in the liquid.
 38. A liquidprocessing apparatus making use of a pipette device comprising a liquidsucking/discharging line which can move in the horizontal direction andis maintained at a specified position so that it can move in thevertical direction; a means for executing liquid sucking/dischargingworks through said liquid sucking/discharging line; a plurality of chipsrequired for processing one type of liquid and provided along thehorizontal direction in which the liquid sucking/discharging line moves;a vessel with said liquid accommodated therein; a magnetic body forattracting magnetic particles contained in a liquid onto an internalsurface of a chip when the liquid is sucked into or discharged from saidchip; and one or more vessels with other types of liquid accommodatedtherein respectively required for the processing above, and the liquidsucking/discharging line or a chip set therein is driven and controlledaccording to instructions from a control unit so that the chip istransferred to execute such works as capturing, extracting, isolating,amplifying, labelling, and measuring a liquid or a target high molecularsubstance contained in the liquid.
 39. A liquid processing apparatusmaking use of a pipette device comprising a liquid sucking/dischargingline which can move in the horizontal direction and is maintained at aspecified position so that it can move in the vertical direction; aplurality of chips required for processing one type of liquid andprovided along the horizontal direction in which the liquidsucking/discharging line moves; a vessel with said liquid accommodatedtherein; one or more filter holders each having a filter required forthe processing above; one or more vessels each accommodating thereinother types of liquid required for the processing above; a vessel inwhich a liquid containing magnetic particles is accommodated; and amagnetic body for attracting said magnetic particles onto an internalsurface of the chip in the process of sucking or discharging a solutioncontaining said magnetic particles, and the liquid sucking/dischargingline is transferred according to instructions from a control unit toautomatically execute such works as quantifying, separating, taking out,pipetting, clearing, condensing, and diluting the liquid or a targethigh molecular substance contained in the liquid and such works asextracting, recovering, and isolating a target high molecular substance.40. A liquid processing apparatus making use of a pipette deviceaccording to any of claim 37 to claim 39, wherein a hook for locking andsupporting a chip engaged in and supported by said liquidsucking/discharging line is rotatably supported by said liquidsucking/discharging line, and said hook is energized in its normal statein the direction in which connection between the liquidsucking/discharging line and the chip is maintained, and also said hookis energized by a lock releasing body provided at a specified positionin the direction in which locking between the liquid sucking/dischargingline and the chip is released.
 41. A liquid processing apparatus makinguse of a pipette device, wherein said filter holder set in a tip sectionof said chip is transferred so that said chip and/or the filter holderis separated from an edge of the liquid sucking/discharging line or achip set therein when the liquid sucking/discharging line locked by thelocking body is raised.
 42. A liquid processing apparatus making use ofa pipette device, wherein a vessel is formed into cassette form having aplurality of chambers each for accommodating a type of liquid thereinand then samples or reagents required for a reaction or processing arepipetted to each of the liquid accommodating chambers, and magneticparticles are absorbed onto an internal surface of said chip by amagnetic force of said magnetic body for transferring.
 43. A liquidprocessing apparatus making use of a pipette device according to claim42, wherein necessary reagents are previously pipetted to each of saidliquid accommodating chambers, and a portion or all of the reagent ineach of said liquid accommodating chambers is shielded with a thin filmbody which can be broken by the liquid sucking/discharging line or achip thereon.
 44. A liquid processing apparatus making use of a pipettedevice according to any of claim 37 to claim 43, wherein said magneticbody is built with a permanent magnet, and a surface of said permanentmagnet contacting a chip is formed according to an external form of thechip and the chip is movably provided in a direction perpendicular tothe longitudinal direction of the chip or in a range including thedirection perpendicular to the longitudinal direction of the chip.
 45. Aliquid processing apparatus making use of a pipette device according toany of claim 37 to claim 43, wherein said magnetic body is built with anelectric magnet, and a surface of said electric magnet contacting a chipis formed according to an external form of the chip, and is provided sothat said electric magnet generates a magnet force when said electricmagnet contacts outside of said chip and also can move, when degaussed,in a direction of separating from the chip.
 46. A liquid processingapparatus making use of a pipette device according to any of claim 44 orclaim 45, wherein a holding body which moves, when said permanent magnetor electric magnet moves to the chip, in synchronism to movement of themagnet is provided in said permanent magnet or electric magnet, asurface of said holding body contacting a chip is formed according to anexternal form of the chip, and the chip is held between said holdingbody and the permanent magnet or electric magnet.
 47. A liquidprocessing apparatus making use of a pipette device, wherein atemperature control step required for a reaction between a target highmolecular substance and a reagent or the like or for amplifying thetarget high molecular substance is inserted into the liquid processingstep with the liquid sucking/discharging line, the reaction liquid orthe liquid to be amplified is transferred with the chip to eachthermostatic vessel kept at a prespecified temperature for controllingthe temperature, and also a covering body, which can be set in a tipsection of the liquid sucking/discharging line, is set by the liquidsucking/discharging line on each thermostatic vessel in which thereaction liquid or the liquid to be amplified is accommodated.
 48. Aliquid processing apparatus making use of a pipette device according toclaim 47, wherein said covering body comprises a flat surface sectionhaving a diameter larger than that of a bore of the thermostatic vesseland a maintenance groove section formed in a substantially centralportion of said flat surface section and having the same bore as anexternal diameter of the liquid sucking/discharging line or the tip ofthe chip, and a bottom section of the maintenance groove section isformed with a thin film body which can be broken by the liquidsucking/discharging line or the chip.